Immunological origins of binding and catalysis in a Diels-Alderase antibody

Science

Volumn 279, Issue 5358, 1998, Pages 1929-1933

  Romesberg, Floyd E ^a,b   Spiller, Ben ^a,b   Schultz, Peter G ^a,b   Stevens, Raymond C ^a,b  

^a UNIVERSITY OF CALIFORNIA   (United States)

^b LAWRENCE BERKELEY NATIONAL LABORATORY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CATALYTIC ANTIBODY; IMMUNOGLOBULIN F(AB) FRAGMENT;

AMINO ACID SUBSTITUTION; ANTIBODY COMBINING SITE; ARTICLE; CATALYSIS; CRYSTAL STRUCTURE; HYDROGEN BOND; PRIORITY JOURNAL; REACTION ANALYSIS;

AMINO ACID SEQUENCE; ANTIBODIES; ANTIBODIES, CATALYTIC; ANTIBODY AFFINITY; ANTIBODY SPECIFICITY; BINDING SITES; BINDING SITES, ANTIBODY; CATALYSIS; CHEMISTRY, ORGANIC; CLONING, MOLECULAR; CRYSTALLOGRAPHY, X-RAY; EVOLUTION, MOLECULAR; GERM-LINE MUTATION; HAPTENS; HYDROGEN BONDING; IMMUNOGLOBULIN FAB FRAGMENTS; MODELS, MOLECULAR; MOLECULAR SEQUENCE DATA; MUTATION; PROTEIN CONFORMATION; RECOMBINANT PROTEINS;

BETULACEAE;

EID: 0032549776     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.279.5358.1929     Document Type: Article

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39. 2+ ions. The germ-line Fab crystallized in the space group C2 with two molecules in the asymmetric unit. The structure was solved similarly with the use of the mature structure as a search model; the final structure contains 868 amino acids and 274 water molecules.
40. 2+ ions. The germ-line Fab crystallized in the space group C2 with two molecules in the asymmetric unit. The structure was solved similarly with the use of the mature structure as a search model; the final structure contains 868 amino acids and 274 water molecules.
41. 2+ ions. The germ-line Fab crystallized in the space group C2 with two molecules in the asymmetric unit. The structure was solved similarly with the use of the mature structure as a search model; the final structure contains 868 amino acids and 274 water molecules.
42. 2+ ions. The germ-line Fab crystallized in the space group C2 with two molecules in the asymmetric unit. The structure was solved similarly with the use of the mature structure as a search model; the final structure contains 868 amino acids and 274 water molecules.
43. Enzyme-linked immunosorbent assay plates were coated with 100 μl of hapten-BSA conjugate (10 μg/ml) in phosphate-buffered saline and incubated overnight at 4°C. Plates were washed, blocked (with phosphate-buffered saline containing 1% BSA and 0.05% Tween 20), and washed again. They were then incubated with 100 μl of 100 nM antibody, washed, and incubated with alkaline phosphatase-conjugated antibodies to human kappa chain. After washing p-nitrophenylphosphate was added to each well and absorbance was measured at 405 nm.
44. Supported by the U.S. Department of Energy, NIH, Howard Hughes Medical Institute (P.G.S.), and W. M. Keck Foundation. We thank Stanford Synchrotron Radiation Laboratory for data collection time. The coordinates have been deposited in the Brookhaven protein database under accession numbers 1A4K (mature) and 1A4J (germ line).